Western Blot Analysis of EMT Pathway

Western blot was carried out according as described [41 (link), 42 (link)] with rabbit polyclonal PLOD2, E-cadherin and β-catenin antibodies (1:1000; Proteintech, USA), PI3K, p-PI3K (Tyr458), AKT, p-AKT (Ser473), GSK-3β, p-GSK-3β, N-Cadherin, Slug and Vimentin antibodies (1:1000; Cell Signaling Technology, Danvers, MA, USA), as well as Snail antibody (1:1000; Abcam, USA). Mouse monoclonal HIF-1α antibody (1:50; Novus Biologicals, USA) was used for normalization. An HRP-conjugated anti-rabbit or anti-mouse IgG antibody was used as the secondary antibody (1:2000; CoWin Bioscience, Beijing, China). Signals were detected using enhanced chemiluminescence reagents (Pierce, Rockford, IL, USA).